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首页> 外文期刊>Journal of Veterinary Diagnostic Investigation >Evaluation of a nested polymerase chain reaction assay to differentiate between two genotypes of Porcine circovirus-2.
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Evaluation of a nested polymerase chain reaction assay to differentiate between two genotypes of Porcine circovirus-2.

机译：评价巢式聚合酶链反应测定以区分猪循环血管孢毛血管孢毛血管孢毛血清血清血清血清血管孢毛血管孢虫-2。

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Porcine circovirus-2 (PCV-2) is associated with several diseases in pigs, including postweaning multisystemic wasting syndrome (PMWS). A new genotype of PCV-2 was isolated from swine farms with and without clinical PMWS in North America. The new genotype was differentiated in a separate cluster by phylogenetic analyses and is now named PCV-2b compared with PCV-2a for the previously known genotype. The purpose of this study was to develop and evaluate a nested polymerase chain reaction (nPCR) assay to detect and differentiate between PCV-2a and PCV-2b. Genotype-specific primer sets were designed by using sequence data published for different PCV-2 strains. Specificity and sensitivity of the nPCR were examined by using PCV-2 isolates with known genotype. Nested PCR was found to be highly specific and sensitive for detecting and differentiating between the PCV-2 genotypes compared with the conventional 1-step PCR assay. Nested PCR was applied to detect PCV-2 and to identify the genotype in serum samples from swine farms with and without a clinical history of PMWS. Of 60 serum samples collected from 4 farms during clinical PMWS outbreaks, PCV-2a and PCV-2b were detected in 6 and 49 samples, respectively. Six of the 10 samples from one of the 4 farms had both PCV-2a and PCV-2b. Of 20 serum samples from 2 farms without PMWS, 11 were positive for PCV-2a only. These results suggest that the differential nPCR can be used to detect PCV-2 and to differentiate the 2 genotypes from field samples.
机译：猪胃肠病毒-2（PCV-2）与猪的几种疾病有关，包括切换多系统浪费综合征（PMW）。从北美的猪场和没有临床PMWS的猪场中分离出一种新的PCV-2基因型。通过系统发育分析，新基因型在单独的簇中分化，并且与先前已知的基因型相比，与PCV-2A相比，现在将PCV-2B命名。本研究的目的是发展和评估嵌套聚合酶链反应（NPCR）测定以检测和分化PCV-2a和PCV-2b。通过使用针对不同PCV-2菌株的序列数据设计基因型特异性引物组。通过使用具有已知基因型的PCV-2分离株检查NPCR的特异性和敏感性。与常规的1步PCR测定相比，发现嵌套的PCR是对PCV-2基因型的检测和分化的高度特异性和敏感性。嵌套的PCR被施用检测PCV-2，并在没有PMW的临床历史上鉴定猪场中血清样品的基因型。在临床PMWS爆发期间从4个农场收集的60个血清样本，分别在6和49个样品中检测到PCV-2A和PCV-2B。来自4个农场之一的10个样品中的六个具有PCV-2A和PCV-2B。来自2个农场的20个血清样品，仅为PCV-2A呈阳性。这些结果表明，差分NPCR可用于检测PCV-2并从场样品中区分2个基因型。

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